AMPK activation induces mitophagy and promotes mitochondrial fission while activating TBK1 in a PINK1-Parkin independent manner

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AMPK activation induces mitophagy and promotes mitochondrial fission while activating TBK1 in a PINK1-Parkin independent manner. / Seabright, Alex; Fine, Nicholas; Barlow, Jonathan; Lord, Samuel; Musa, Ibrahim; Gray, Alexander; Bryant, Jack; Banzhaf, Manuel; Lavery, Gareth; Hardie, David Grahame; Hodson, David; Philp, Andrew; Lai, Yu-Chiang.

In: FASEB Journal, Vol. 34, No. 5, 05.2020, p. 6284-6301.

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Seabright, Alex ; Fine, Nicholas ; Barlow, Jonathan ; Lord, Samuel ; Musa, Ibrahim ; Gray, Alexander ; Bryant, Jack ; Banzhaf, Manuel ; Lavery, Gareth ; Hardie, David Grahame ; Hodson, David ; Philp, Andrew ; Lai, Yu-Chiang. / AMPK activation induces mitophagy and promotes mitochondrial fission while activating TBK1 in a PINK1-Parkin independent manner. In: FASEB Journal. 2020 ; Vol. 34, No. 5. pp. 6284-6301.

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@article{2cb3b280d64a45bda19fb24ac2759364,
title = "AMPK activation induces mitophagy and promotes mitochondrial fission while activating TBK1 in a PINK1-Parkin independent manner",
abstract = "Mitophagy is a key process regulating mitochondrial quality control. Several mechanisms have been proposed to regulate mitophagy, but these have mostly been studied using stably expressed non-native proteins in immortalized cell lines. In skeletal muscle, mitophagy and its molecular mechanisms require more thorough investigation. To measure mitophagy directly, we generated a stable skeletal muscle C2C12 cell line, expressing a mitophagy reporter construct (mCherry-green fluorescence protein-mtFIS1 101-152). Here, we report that both carbonyl cyanide m-chlorophenyl hydrazone (CCCP) treatment and adenosine monophosphate activated protein kinase (AMPK) activation by 991 promote mitochondrial fission via phosphorylation of MFF and induce mitophagy by ~20%. Upon CCCP treatment, but not 991, ubiquitin phosphorylation, a read-out of PTEN-induced kinase 1 (PINK1) activity, and Parkin E3 ligase activity toward CDGSH iron sulfur domain 1 (CISD1) were increased. Although the PINK1-Parkin signaling pathway is active in response to CCCP treatment, we observed no change in markers of mitochondrial protein content. Interestingly, our data shows that TANK-binding kinase 1 (TBK1) phosphorylation is increased after both CCCP and 991 treatments, suggesting TBK1 activation to be independent of both PINK1 and Parkin. Finally, we confirmed in non-muscle cell lines that TBK1 phosphorylation occurs in the absence of PINK1 and is regulated by AMPK-dependent signaling. Thus, AMPK activation promotes mitophagy by enhancing mitochondrial fission (via MFF phosphorylation) and autophagosomal engulfment (via TBK1 activation) in a PINK1-Parkin independent manner. ",
keywords = "endogenous, mitophagy, skeletal muscle, tandem ubiquitin-binding entity (TUBE), ubiquitin",
author = "Alex Seabright and Nicholas Fine and Jonathan Barlow and Samuel Lord and Ibrahim Musa and Alexander Gray and Jack Bryant and Manuel Banzhaf and Gareth Lavery and Hardie, {David Grahame} and David Hodson and Andrew Philp and Yu-Chiang Lai",
note = "{\textcopyright} 2020 The Authors. The FASEB Journal published by Wiley Periodicals, Inc. on behalf of Federation of American Societies for Experimental Biology.",
year = "2020",
month = may,
doi = "10.1096/fj.201903051R",
language = "English",
volume = "34",
pages = "6284--6301",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "Federation of American Society of Experimental Biology",
number = "5",

}

RIS

TY - JOUR

T1 - AMPK activation induces mitophagy and promotes mitochondrial fission while activating TBK1 in a PINK1-Parkin independent manner

AU - Seabright, Alex

AU - Fine, Nicholas

AU - Barlow, Jonathan

AU - Lord, Samuel

AU - Musa, Ibrahim

AU - Gray, Alexander

AU - Bryant, Jack

AU - Banzhaf, Manuel

AU - Lavery, Gareth

AU - Hardie, David Grahame

AU - Hodson, David

AU - Philp, Andrew

AU - Lai, Yu-Chiang

N1 - © 2020 The Authors. The FASEB Journal published by Wiley Periodicals, Inc. on behalf of Federation of American Societies for Experimental Biology.

PY - 2020/5

Y1 - 2020/5

N2 - Mitophagy is a key process regulating mitochondrial quality control. Several mechanisms have been proposed to regulate mitophagy, but these have mostly been studied using stably expressed non-native proteins in immortalized cell lines. In skeletal muscle, mitophagy and its molecular mechanisms require more thorough investigation. To measure mitophagy directly, we generated a stable skeletal muscle C2C12 cell line, expressing a mitophagy reporter construct (mCherry-green fluorescence protein-mtFIS1 101-152). Here, we report that both carbonyl cyanide m-chlorophenyl hydrazone (CCCP) treatment and adenosine monophosphate activated protein kinase (AMPK) activation by 991 promote mitochondrial fission via phosphorylation of MFF and induce mitophagy by ~20%. Upon CCCP treatment, but not 991, ubiquitin phosphorylation, a read-out of PTEN-induced kinase 1 (PINK1) activity, and Parkin E3 ligase activity toward CDGSH iron sulfur domain 1 (CISD1) were increased. Although the PINK1-Parkin signaling pathway is active in response to CCCP treatment, we observed no change in markers of mitochondrial protein content. Interestingly, our data shows that TANK-binding kinase 1 (TBK1) phosphorylation is increased after both CCCP and 991 treatments, suggesting TBK1 activation to be independent of both PINK1 and Parkin. Finally, we confirmed in non-muscle cell lines that TBK1 phosphorylation occurs in the absence of PINK1 and is regulated by AMPK-dependent signaling. Thus, AMPK activation promotes mitophagy by enhancing mitochondrial fission (via MFF phosphorylation) and autophagosomal engulfment (via TBK1 activation) in a PINK1-Parkin independent manner.

AB - Mitophagy is a key process regulating mitochondrial quality control. Several mechanisms have been proposed to regulate mitophagy, but these have mostly been studied using stably expressed non-native proteins in immortalized cell lines. In skeletal muscle, mitophagy and its molecular mechanisms require more thorough investigation. To measure mitophagy directly, we generated a stable skeletal muscle C2C12 cell line, expressing a mitophagy reporter construct (mCherry-green fluorescence protein-mtFIS1 101-152). Here, we report that both carbonyl cyanide m-chlorophenyl hydrazone (CCCP) treatment and adenosine monophosphate activated protein kinase (AMPK) activation by 991 promote mitochondrial fission via phosphorylation of MFF and induce mitophagy by ~20%. Upon CCCP treatment, but not 991, ubiquitin phosphorylation, a read-out of PTEN-induced kinase 1 (PINK1) activity, and Parkin E3 ligase activity toward CDGSH iron sulfur domain 1 (CISD1) were increased. Although the PINK1-Parkin signaling pathway is active in response to CCCP treatment, we observed no change in markers of mitochondrial protein content. Interestingly, our data shows that TANK-binding kinase 1 (TBK1) phosphorylation is increased after both CCCP and 991 treatments, suggesting TBK1 activation to be independent of both PINK1 and Parkin. Finally, we confirmed in non-muscle cell lines that TBK1 phosphorylation occurs in the absence of PINK1 and is regulated by AMPK-dependent signaling. Thus, AMPK activation promotes mitophagy by enhancing mitochondrial fission (via MFF phosphorylation) and autophagosomal engulfment (via TBK1 activation) in a PINK1-Parkin independent manner.

KW - endogenous

KW - mitophagy

KW - skeletal muscle

KW - tandem ubiquitin-binding entity (TUBE)

KW - ubiquitin

UR - http://www.scopus.com/inward/record.url?scp=85082034250&partnerID=8YFLogxK

U2 - 10.1096/fj.201903051R

DO - 10.1096/fj.201903051R

M3 - Article

C2 - 32201986

VL - 34

SP - 6284

EP - 6301

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 5

ER -